MRSA is one of the major pathogens in hospitals and the community, which have the ability to produce biofilm as a virulence factor, the impact of chalcone on biofilm formation, the synergism effect of chalcone and antibiotic in both in vitro and in vivo experiments, the gene expression of virulence genes (srtA, fnbA, fnbB) before and after treatment of it on MRSA biofilm cells in vitro, all these were the prime aims of this study. Chalcone at MBIC (20 μg/ml), significantly reduced the biofilm formation to 21.45% and at sub MBIC (15 μg/ml) to 36.58 %. While, Chalcone at MIC(5 μg/ml) reduced MRSA planktonic cells to 49.61%. Susceptibility of MRSA isolates against eight antibiotics showed that all isolates were sensitive to vancomycin and none of the isolates developed susceptibility to erythromycin. The combinatorial effect of chalcone at 5 μg/ml and vancomycin at MIC of (1 μg/ml) on MRSA planktonic cells was reduced it from 70 to 23.3% , and in combination with erythromycin at 32 μg/ml, was decreased from 53.1% to 22% and the effect of chalcone at sub MBIC (15 μg/ml) when combined with vancomycin was reducing the biofilm formation from 87% to 27.6 and with erythromycin from 55.1% to 23.8%. Combinatorial phenotypic effect of the antibiotics and chalcone (at sub MBIC), in vitro came in line with the result of in vivo experiment and the results showed decrease in the expression of fnbA, fnbB and srtA genes in tested isolates in the presence of chalcone at sub MBIC. In our study, we demonstrated that chalcone exhibited significant effect in biofilm formation of MRSA strains, which can be considered as promising antimicrobial agents that can be used for prevention of MRSA adherence or as adjunct to antibiotics in conventional therapy.
Normally, bacteria exposed to antibiotics at sub minimal inhibitory concentrations (MIC) inside the host. Therefore, the current study aimed to comprehend the association among hemolysins, biofilm, as well as gentamicin resistance in local MRSA isolates. Around 35 Staphylococcus aureus locally isolated from different clinical specimens were employed in this study. Methicillin resistance was detected via cefoxitin disk diffusion and mecA amplification methods. MIC of gentamicin was estimated by broth microdilution method. Hemolysin genes involving hla, hlb, hld, and hlg were determined using multiplex polymerase chain reaction (PCR) technique. Microtiter plate method was employed for biofilm assessment in the presence and absence of gentamic
... Show MoreStaphylococcus aureus and Pseudomonas aeruginosa are the major globally distributed pathogens, which causes chronic and recalcitrant infections due to their capacity to produce biofilms in large part. Biofilm production represents a survival strategy in these species, allowing them to endure environmental stress by altering their gene expression to match their own survival needs. In this study, we co-cultured different clinical isolates of S. aureus and P. aeruginosa as mono- and mixed-species biofilms in a full-strength Brain Heart Infusion Broth (BHI) and in a 1000-fold diluted Brain Heart Infusion Broth (BHI/1000) using Microtiter plate assay and determination of colony-forming units. Furthermore, the effect of starvation stress on the e
... Show MoreThe resistance of Staphylococcus aureus to ciprofloxacin has complicated the problem of treating staphylococcal associated infections in which MRSA is the causative agent since ciprofloxacin was the drug of choice to treat such infections. Our study investigated the incidence of Ciprofloxacin resistant S. aureus isolates that were also methicillin resistant among Iraqi patients. The obtained bacterial isolates were tested for Ciprofloxacin resistance using agar dilution method and the sequence of gyrA and parC. The results revealed that about 8% of the isolated MRSA strains were Ciprofloxacin resistant and the resistance was due to mutation in gyrA rather than parC.
Rapid and accurate identification of Methicillin Resistant Staphylococcus aureus is essential in limiting the spread of this bacterium. The aim of study is the detection of Methicillin Resistant Staphylococcus aureus (MRSA) and determining their susceptibility to some antimicrobial agent. A total of fifty clinical Staphylococcus aureus, isolated from the nose of health work staff in surgery unit of Kalar general hospital and from ear of patients attended to the same hospital. The susceptibilities of isolates were determined by the disc diffusion method with oxacillin (1 ?g) and cefoxitin (30 ?g), and by the mannitol salt agar supplemented with cefoxitin (MSA-CFOX), susceptibilities of isolates to other antimicrobial agent were determined b
... Show MoreObjective: The present work was undertaken to investigate the impact of sub inhibitory concentration of gentamicin on hla gene expression in methicillin resistant Staphylococcus aureus isolates. Methods: The bacterial isolates used in this study represent 33 MRSA strains, previously isolated form patients visiting several hospitals in Baghdad. Gentamicin, vancomycin, and oxacillin MIC were determined using broth dilution method. Microtiter plate method was adopted to investigate the biofilm forming capacity. Alpha hemolysin was detected by culturing MRSA isolates on rabbit blood agar. Furthermore, hla gene was detected in MRSA isolates using conventional PCR technique; while, qRT-PCR method was performed to assay the hla expression in plank
... Show MorePresent study was carried out to find prevalence of MRSA in healthy individual of second stage students, college of pharmacy/Baghdad University. A total of 74 student selected between age 18-23 years old were included in this study, nasal swabs collected and subjected to many diagnostic standard bacteriological identification methods. Culture, colonial morphology, Gram stain, mannitol fermentation, coagulase ,gelatinasetest, DNAase, MR/VP and antimicrobial susceptibility test was performed on tryptic soy agar by modified Kirby-Bauer muller hinton disc diffusion method and the result show that out of 74 nasal swabs,67(90.5%) were MRSA positive isolates, 21(31.4%) of them were mannitol ferment and 46(68.6%) non mannitol fermenter, am
... Show MoreOut of 150 different specimens, 67 S. aureus isolate were isolated. However, 16sRNA gene was located only in 60 isolates. Moreover, mecA gene was located in 48 isolates; thereby MRSA covered 80% of all S. aureus isolates. Of considerable interest, pvl gene was detected in only six isolates (10%). Hence, the present work emphasizes the notion suggested that pvl is not an indicative of CA-MRSA.
The photodynamic inactivation against Methicillin-resistant Staphylococcus aureus using two different lasers, 532 nm diode pumped solid state laser (DPSS) in combination with safranin O and 650 nm diode laser in combination with methylene blue was investigated in the present work. A hundred swab samples were collected from patients with burn and wound infections admitted to two hospitals in Baghdad (Specialized Burns Hospital in Medical City and Al Imamein Al Jwadein Medical City Hospital) from December 2015 to February 2016 Antimicrobial susceptibility was performed by using Kirby- Bauer method. The irradiation experiments included four groups; a control group, a photosensitizer only group, a laser irradiation only group and a laser irr
... Show MoreScrophularia. striata from Scrophulariacea family has been used in Iranian folk medicine for the treatment of infectious diseases. In this study we evaluated the synergistic effect of S. striata hydroalcoholic extract (SSE) and commercially available antibiotics against P. aeroginosa and Methicillin- resistant Staphylococcus aureus (MRSA). The resazurin-based microdilution method was used to determine the minimum inhibitory concentration (MIC) values of plan extract and standard antibiotics. The interaction between standard antibiotics and SSE was evaluated by using checkerboard method. The results of this study revealed that SSE enhance the antibacterial activity of antibiotics. The combin
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